Synergistic composition for maintenance of healthy balance of microflora

ABSTRACT

The present invention relates to compositions, particularly compositions useful in maintaining and supporting healthy microflora in the female urogenital tract which could lead to inhibition of vaginal infections, as well as methods of treating and preventing vaginal infections. Compositions useful in supporting healthy microflora, disclosed herein, generally comprise a therapeutic amount of a first saccharide and a therapeutic amount of a second saccharide or an organic acid. The saccharides may be, for example, a pentose, a disaccharide, a cyclodextrin, a pectic substance or a non-digestible polysaccharide. The organic acid may be, for example, malic acid.

BACKGROUND OF THE DISCLOSURE

Humans are colonized by microbes in the gastrointestinal tract, on theskin, and in other epithelial and tissue niches such as the oral cavity,eye surface and vagina. In healthy persons a single local or tissue typemay be inhabited by hundreds of different species of bacteria.Interactions between various bacteria species in these populations andbetween bacteria and the human host shape the community structure withavailability of and competition for resources affecting the distributionof various species of bacteria. Such resources may be food, location andthe availability of space to grow or a physical structure to which thebacteria may attach.

A healthy microbial flora provides the host with multiple benefits,including colonization resistance to a broad spectrum of pathogens,essential nutrient biosynthesis and absorption, and immune stimulation.For example, a normal vagina generally contains more than about 10⁴lactobacilli per milliliter of vaginal fluid. Under normal conditions,the vagina flora provides a mildly acidic environment that helps guardagainst the invasion of pathogenic microbes. Unfortunately, this vaginalbalance may be easily upset by a variety of external factors thatultimately lead to vaginal infection. Vaginal infection is a clinicalsyndrome and exists in three primary forms, i.e., bacterial vaginosis,candidal vaginitis (“yeast”), and trichomonas vaginitis (“trich”).

Current treatment regimens for bacterial infection of the vagina involvethe use of various broad spectrum antibiotics, such as metronidazole.However, antibiotics are often undesirable because they may kill a broadrange of the normal bacterial flora in the vagina, including thebeneficial lactobacilli. This may cause secondary complications, becausethe lactobacilli keep various opportunistic pathogens in the vagina incheck. The treatment may then necessitate a further treatment regimen,such as the ingestion of cultured dairy products to replace thelactobacilli in the body, as well as treatment by antifungal agents.Moreover, a rise in the level of anaerobes due to a lack of lactobacillicould further complicate the infection. Additionally, antibiotics, whenused frequently within the vagina, may cause systemic toxicity throughabsorption from the vagina.

As such, a need currently exists for improved compositions forsupporting and maintaining a healthy balance of microflora in theurogenital area and more particularly improved vaginal treatmentcompositions.

SUMMARY OF THE DISCLOSURE

It has now been surprisingly discovered that the growth of certainstrains of Lactobacilli may be synergistically increased byadministering a composition comprising two different carbon sources. Thecarbon sources may consist of a first saccharide and a second saccharideor an organic acid. Increasing the growth of beneficial lactobacilli mayeffectively inhibit the growth of pathogens associated with urogenitalinfections and help maintain a healthy microflora balance in theurogenital area. As such compositions comprising a first saccharide anda second saccharide or an organic acid are well suited for topicaladministration to the urogenital area of a female for support andmaintain a healthy balance of microflora in the urogenital area. Forexample, providing a composition comprising a pentose and a disaccharidesynergistically promotes the growth of Lactobacillus spp. withoutpromoting growth of pathogenic bacteria such as Escherichia coli orGardnerella vaginalis. Accordingly, in one embodiment the presentinvention provides a composition comprising a first therapeutic agentconsisting of a pentose or a disaccharide and a second therapeutic agentselected from the group consisting of a non-digestible polysaccharide,an organic acid, a cyclodextrin, a pectic substance, a pentose or adisaccharide.

In other embodiments the present invention provides a compositioncomprising a disaccharide selected from the group consisting oflactulose, trehalose, rhamnose maltose, maltotriose, lactose andlactitol and a pentose selected from the group consisting of ribose,ribulose, arabinose, xylose, xylulose, and lyxose, methylβ-D-ribofuranoside and 2-deoxy-D-ribose. In a particularly preferredembodiment the disaccharide comprises from about 0.05 to about 1.0wt/vol % and the ratio of disaccharide to pentose is from about 1:1 toabout 1:10.

In still other embodiments the present invention provides a compositioncomprising a pentose selected from the group consisting of ribose,ribulose, arabinose, xylose, xylulose, and lyxose, methylβ-D-ribofuranoside and 2-deoxy-D-ribose and an organic acid selectedfrom the group consisting of citric acid, lactic acid, methyllacticacid, phenyllactic acid, malic acid, mandelic acid, glycolic acid,tartronic acid, tartaric acid. In a particularly preferred embodimentthe pentose comprises from about 0.05 to about 1.0 wt/vol % and theratio of pentose to organic acid is from about 1:1 to about 1:10.

In yet other embodiments the present invention provides a compositioncomprising a first therapeutic agent selected from the group consistingof α-methyl-D-glucoside, β-methyl-D-glucopyranoside and salicin and asecond therapeutic agent selected from the group consisting of apentose, a disaccharide, a non-digestible polysaccharide, an organicacid, a cyclodextrin and a pectic substance.

In other embodiments the present invention provides compositions foradministration to a user. Suitable formulations may include, forexample, liquids, solutions, pastes or gels. Accordingly, in onepreferred embodiment the present invention provides a formulationcomprising a first therapeutic agent consisting of a pentose or adisaccharide and a second therapeutic agent consisting of an organicacid, a cyclodextrin, a pectic substance, a pentose or a disaccharideand a from about 0.05 to about 5.0 wt/vol of at least one gelling agentthat includes gellan gum. In a particularly preferred embodiment thefirst therapeutic agent and the second therapeutic agent comprise fromabout 0.1 to about 2.0 wt/vol % and the ratio of the first therapeuticagent to the second therapeutic agent is from about 1:1 to about 1:10.In a particularly preferred embodiment the formulations may be topicallyapplied to the urogenital area and are capable of supporting andmaintaining a healthy balance of microflora in the urogenital area.

In yet other embodiments the compositions of the present invention maybe applied to an applicator. Suitable applicators include a web, such asa wet laid tissue web or air laid web, gauze, cotton swab, transdermalpatch, container or holder. Thus, in certain embodiments the compositionmay be applied to a nonwoven web, such as meltblown, coform, spunbond,airlaid, hydroentangled nonwovens, spunlace, bonded carded webs, andlaminates thereof, as well as wet laid fibrous webs, such as tissuewebs. Accordingly, in one embodiment the invention provides atherapeutic wipe comprising a nonwoven web and a composition disposedthereon, the composition comprising a pentose and a disaccharide.

In other aspects the compositions of the present invention may beadministered to a user to maintain and support growth of Lactobacillusspp. and inhibit pathogens. Accordingly, in one embodiment the presentinvention provides a method for enhancing lactobacillus growth oractivity in vivo comprising administering a composition comprising afirst therapeutic agent consisting of a pentose or a disaccharide and asecond therapeutic agent consisting of an organic acid, a cyclodextrin,a pectic substance, a pentose or a disaccharide.

Definitions

As used herein, the term “inhibit” generally means to reduce by ameasurable amount or to prevent entirely.

As used herein the term “urogenital” refers to the vulva, vagina,urinary tract, bladder, and surrounding areas.

As used herein the terms “effective amount” and “therapeutic amount” isan amount sufficient to maintain and support a healthy balance ofmicroflora. In fact, although not required, it may be desired to use aconcentration that does not significantly affect or inhibit the growthcharacteristics of the normal vaginal flora or otherwise significantlyirritate the vaginal tissue when used at inhibitory, noncytotoxic, orclinical concentrations. For example, the therapeutic agent(s) aredesirably employed at a concentration of about 0.01 to about 20.0 wt/vol%, in some embodiments from about 0.1 wt/vol % to about 10.0 wt/vol %,in some embodiments from about 0.2 to about 5.0 wt/vol %, and in someembodiments from about 0.5 to about 4.5 wt/vol %. It should beunderstood that the dosage may vary with the age, condition, and type ofinfection suffered by the patient, and may be readily determined by oneof skill in the art.

As used herein the term “therapeutic effect” refers to the ability ofthe compositions and formulations of the present invention to stimulatethe growth of L. crispatus relative E. coli measured according to thetherapeutic effect protocol described below. Generally therapeuticeffect is expressed as a ratio of L. crispatus to E. coli and isdesirably greater than about 30, more preferably greater than about 50and more desirably greater than about 100.

As used herein, the designation “wt/vol %” or “wt/vol” refers to thevalue obtained by dividing the weight of a substance (in grams) by thevolume of the solution (in milliliters), and then multiplying by 100.

As used herein the term “non-digestible polysaccharide” as used in thepresent invention refers to polysaccharides which are not or onlypartially digested in the intestine by the action of acids or digestiveenzymes present in the human upper digestive tract (small intestine andstomach) but which are fermented by the human intestinal flora. Forexample, sucrose, lactose, maltose and maltodextrins are considereddigestible. Preferably the non-digestible polysaccharide is anon-digestible neutral polysaccharide wherein more than 75% of thesaccharides units are selected from the group consisting of glucose,fructose, galactose, mannose, ribose, rhamnose, arabinose, and xylose,preferably more than 85%, more preferably more than 95%, even morepreferably more than 99%. Preferably the present non-digestiblepolysaccharide is a prebiotic polysaccharide that stimulates the growthand/or activity of one or a limited number of probiotic bacterialspecies in the colon.

As used herein the term “pentose” generally refers to a monosaccharidecomprising a five-membered furanose ring. Suitable pentoses may have thegeneral formula C₅H₁₀O₅ such as, for example, ribose, ribulose,arabinose, xylose, xylulose, and lyxose, and isomers thereof. The termpentose also includes five-membered furanose rings reacted under acidconditions to form an acetal such as methyl δ-D-ribofuranoside. The termpentose also includes five-membered furanose rings derived from apentose having the general formula C₅H₁₀O₅ by the loss of an oxygen atomsuch as 2-deoxy-D-ribose. Pentoses are preferably provided in the formof a five-membered furanose ring and therefore do not include sugaralcohols, which may have the same linear structure as pentoses, but aremodified with one or more alcohol groups.

As used herein the term “pectic substance” generally refers to pectins,pectates, polygalacturonic acids, and mixtures thereof. Pectic substanceare generally complex organic polysaccharides derived from the secondarycell wall of terrestrial plants. The polygalacturonic acid referred toherein is composed of repeating units of the galacturonic acid units arejoined by α-(1→4) linkages. Pectin contains as its major componentgalacturonic methyl ester which is the methyl ester of a galacturan.However the galacturonic acid units are not fully esterified. Thegalacturan ethyl esters are linear molecules with molecular weights ofapproximately 30,000 to about 300,000.

As used herein the term “saccharide” generally refers to apolysaccharide, an oligosaccharide, or a monosaccharide. Frequently,references to a saccharide refers to a monosaccharide, such as apentose, a disaccharide, such as lactulose, trehalose, rhamnose maltose,maltotriose, lactose and lactitol, a cyclodextrin, pectin, or anon-digestible polysaccharide.

As used herein the term “soluble” when having reference to a pentose, adisaccharide, an organic acid, a cyclodextrin, a pectic substance or anon-digestible polysaccharide means that the substance is at leastsoluble according to the method described by L. Prosky et al, J. Assoc.Off. Anal. Chem. 71, 1017-1023 (1988).

DETAILED DESCRIPTION OF THE DISCLOSURE

The present invention is related to compositions useful in maintainingand supporting healthy microflora. The compositions are particularlywell suited for administration to the urogenital tract to support andmaintain a healthy microflora. Additionally the compositions andformulations of the present invention may be useful in supporting andmaintaining healthy microflora balance on skin, in the bladder or thegastro-intestinal tract. For example, maintenance and support of ahealthy microflora may be achieved by topically administering acomposition to the urogenital tract or other area of the body. In otherembodiments the compositions of the present invention may be formulatedfor oral administration and orally administered to a patient to supportand maintain healthy microflora in the gastro-intestinal tract.

Compositions useful in supporting and maintaining healthy microfloragenerally comprise a therapeutic amount of a first therapeutic agent anda therapeutic amount of a second therapeutic agent which may be asaccharide or an organic acid. The first therapeutic agent may beα-methyl-d-glucoside, β-methyl-D-glucopyranoside or salicin. In otherembodiments the first therapeutic agent may be selected from the groupconsisting of a pentose, a disaccharide, a non-digestiblepolysaccharide, an organic acid, a cyclodextrin and a pectic substance.

Saccharides useful as therapeutic agents may be, for example, a pentose,a disaccharide, a cyclodextrin, a pectic substance or a non-digestiblepolysaccharide. Organic acids useful as therapeutic agents may be, forexample, citric acid, lactic acid, methyllactic acid, phenyllactic acid,malic acid, mandelic acid, glycolic acid, tartronic acid, tartaric acidand gluconic acid. A particularly preferred organic acid is malic acid.

Urogenital treatment compositions of the present invention generallystimulate the growth of healthy, native, bacteria such as Lactobacillusspp. and may be administered in several forms to a user. For example,the urogenital compositions may be prepared as formulations foradministration to a user or may be applied to a substrate, such as awiping substrate, for administration to a user. Preferably thesaccharides useful in the present invention are soluble to facilitatetheir formulation for administration to a user.

Surprisingly compositions comprising a first saccharide and a secondsaccharide or an organic acid synergistically promote the growth ofhealthy bacteria such as Lactobacillus spp. and more particularlyLactobacillus crispatus without promoting growth of pathogenic bacteria,such as Gardnerella (e.g., Gardnerella vaginalis) or Candida (e.g.,Candida albicans), Accordingly, compositions of the present inventionmay be administered to a user to synergistically and selectivelystimulate growth of lactobacilli without stimulating the growth ofcompeting pathogenic bacteria. Thus, in-use, administration of aformulation comprising a first saccharide and a second saccharide or anorganic acid may enhance the growth and colonization of healthy bacteriasuch as Lactobacillus spp. in the user, which thereby helps reduce theincidence of disease.

Accordingly, in a preferred embodiment, the urogenital compositioncomprises a first therapeutic agent and a second therapeutic agentwherein the composition synergistically effects the growth of L.crispatus over E. coli as measured using the therapeutic effect protocoldescribed below. Preferably the composition yields a ratio of L.crispatus to E. coli greater than about 30, still more preferablygreater than about 50 and still more preferably greater than 200, andeven more preferably greater than about 300.

Generally compositions useful in the present invention comprise at leasttwo therapeutic agents. For example, the first therapeutic agent may beα-methyl-d-glucoside, β-methyl-D-glucopyranoside or salicin and thesecond therapeutic agent may be selected from the group consisting of apentose, a disaccharide, a non-digestible polysaccharide, an organicacid, a cyclodextrin and a pectic substance. Preferably the first andsecond therapeutic agents are different.

In certain embodiments the first therapeutic agent may comprise asaccharide and the second therapeutic agent may be an organic acid or asaccharide. For example, in one embodiment, the composition may comprisea pentose in combination with a disaccharide. Suitable pentoses mayinclude ribose, ribulose, arabinose, xylose, xylulose, and lyxose,methyl β-D-ribofuranoside and 2-deoxy-D-ribose. Particularly preferredpentoses include arabinose and 2-deoxy-D-ribose. Preferably the pentoseshave not been modified with one or more alcohol groups. Suitabledisaccharides may be selected from the group consisting of lactulose,trehalose, rhamnose maltose, maltotriose, lactose and lactitol.Particularly preferred disaccharides are lactulose and trehalose.Accordingly, in one preferred embodiment the composition comprisesarabinose or 2-deoxy-D-ribose and lactulose or trehalose. In otherpreferred embodiments the composition comprises lactulose or trehaloseand a second therapeutic agent selected from the groups consisting of apectic substance, a cyclodextrin, a non-digestible polysaccharides, apentose and an organic acid.

In other embodiments the one of the therapeutic agents may be a pecticsubstance selected from the group consisting of pectins, pectates, andpolygalacturonic acids. The pectic substance is preferably combined witha pentose, a disaccharide or a cyclodextrin to produce a compositionuseful in the present invention. Thus, in a particularly preferredembodiment the composition comprises a pectic substance and a pentoseselected from the group consisting of ribose, arabinose,2-deoxy-D-ribose and methyl β-D-ribofuranoside. In other embodiments thecomposition comprises a pectic substance and a disaccharide selectedfrom the group consisting of lactulose, trehalose, rhamnose maltose,maltotriose, lactose and lactitol. In particularly preferred embodimentsthe composition comprises a first therapeutic component consisting oftrehalose, lactulose, arabinose, or 2-deoxy-D-ribose and a secondtherapeutic component consisting of pectin.

In still other embodiments the therapeutic agent may be an organic acid.Organic acids useful in the present invention generally consist of mono-or polycarboxylic acids having one or more hydroxyl functional groups atleast one of which is introduced into the α-position (i.e., on thecarbon atom adjacent to the carboxyl functional group). Examples ofparticularly useful organic acids include citric acid, lactic acid,methyllactic acid, phenyllactic acid, malic acid, mandelic acid,glycolic acid, tartronic acid, tartaric acid and gluconic acid. Inparticularly preferred embodiments the organic acid is selected from thegroup consisting of citric acid, lactic acid, malic acid, glycolic acidand tartaric acid. In certain embodiments the organic acid may beprovided with an appropriate counterion, such as calcium, sodium ormagnesium. In particularly preferred embodiments the compositioncomprises a first therapeutic component consisting of trehalose,lactulose, arbinose, or 2-deoxy-D-ribose and a second therapeuticcomponent consisting of an organic acid. In other preferred embodimentsthe composition comprises a first therapeutic agent and an organic acidselected from the group consisting of citric acid, lactic acid, malicacid, glycolic acid and tartaric acid, where the first therapeutic agentis trehalose, lactulose, arabinose, or 2-deoxy-D-ribose.

In other embodiments the therapeutic agent may be a cyclodextrin.Suitable cyclodextrins useful in the present invention includehydroxypropyl β-cyclodextrin, hydroxyethyl β-cyclodextrin, hydroxypropylγ-cyclodextrin, hydroxyethyl γ-cyclodextrin, α-cyclodextrin and methylβ-cyclodextrin. Suitable cyclodextrins typically have to have an aqueoussolubility of at least about 10% by weight. Alpha-cyclodextrin is apreferred cyclodextrin. In one embodiment the composition comprises afirst therapeutic component selected from the group consisting of apentose, a disaccharide or a pectic substance and α-cyclodextrin.

In still other embodiments the therapeutic agent may be a polysaccharidethat is not digestible by humans. Suitable non-digestiblepolysaccharides include, for example, dextrin, inulin,fructo-oligosaccharide (FOS) and isomalto-oligosaccharides. In aparticularly preferred embodiment the non-digestible polysaccharidecomprises at least one beta-glycosidic (e.g., beta galactosidic or betaglucosidic) bond or at least one α-glycosidic (e.g., α-galactosidic orα-glucosidic) bond, and is non-digestible by a human digestive system,but can be digested by a bacterium. In one embodiment the bacterium is aBifidobacterium spp. or Lactobacillus spp. Particularly preferrednon-digestible polysaccharides include dextrin and inulin. In aparticularly preferred embodiment the composition comprises dextrin orinulin and a second therapeutic agent selected from the group consistingof a pentose, a disaccharide, cyclodextrin and a pectic substance.

Generally compositions of the present invention comprise less than about10.0 wt/vol % therapeutic agent. That is to say, that the total amountall therapeutic agents, such as pentose, disaccharide, pectic substance,cyclodextrin and organic acid, is generally less than about 10.0 wt/vol%. In particularly preferred embodiments the total amount of therapeuticagent is less than about 5.0 wt/vol % and still more preferably lessthan about 2.5 wt/vol %, such as from about 0.1 to about 2.0 wt/vol %percent and more preferably from about 0.2 to about 1.5 wt/vol %. Forexample, in one embodiment, the composition comprises from about 0.1 toabout 2.0 wt/vol % disaccharide selected from the group consisting ofselected from the group consisting of lactulose, trehalose, rhamnosemaltose, maltotriose, lactose and lactitol and from about 0.1 to about2.0 wt/vol % of a pentose, a pectic substance, a cyclodextrin or anorganic acid.

Further, the first and second therapeutic agents should be provided inan amount sufficient to provide a synergistic effect when administeredto a user. For example, where the composition comprises a pentose and anorganic acid the pentose and organic acid are present in an amountsufficient to stimulate the growth of certain healthy bacteria such asLactobacillus bulgaricus, Lactobacillus acidophilus, Lactobacillusgasseri, Lactobacillus crispatus, Lactobacillus casei, Lactobacillusplantarum, Streptococcus faecium, and Streptococcus thermophilus.Accordingly, in certain embodiments, the organic acid may range from0.05 to 2.0 wt/vol %, such as from about 0.1 to about 1.5 wt/vol % andmore preferably from 0.1 to 1.0 wt/vol % and the pentose may range fromabout 0.1 to about 2.0 wt/vol % and more preferably from about 0.5 toabout 1.5 wt/vol %.

Generally compositions of the present invention comprise two differenttherapeutic agents and more preferably therapeutic agents havingdistinct molecular compositions. That is to say, that where onetherapeutic agent is a disaccharide the second therapeutic agent is nota disaccharide or where one therapeutic agent is a pentose the secondtherapeutic agent is not a pentose. Particularly preferred combinationsof therapeutic agents include, for example, a disaccharide and pentose,a disaccharide and a cyclodextrin, a disaccharide and a pecticsubstance, a pentose and a pectic substance, a pentose and an organicacid, a pectic substance and a non-digestible polysaccharide, a pentoseor a disaccharide and a non-digestible polysaccharide, and a pentose anda cyclodextrin.

The amount of the first and second therapeutic agents may be variedrelative to one another. For example, in certain embodiments, the ratioof the first therapeutic agent to the second therapeutic agent may rangefrom about 1:1 to about 1:50, such as from about 1:1 to about 1:20 andmore preferably from about 1:1 to about 1:5. Accordingly, in certainembodiments the composition may comprises from about 0.1 to about 2.0wt/vol % disaccharide or pentose and from about 0.1 to about 1.0 wt/vol% organic acid, pectic substance or cyclodextrin.

The compositions of the present invention may be formulated foradministration to a user. The composition is generally applied in theform of a douche formulation, spray, moisturizer, lotion, cream, jelly,liniment, ointment, salve, oil, foam, gel, film, wash, suppository,slow-releasing polymer, coating, liquid, vaginal capsule, vaginaltablet, vaginal film, vaginal sponge, vaginal ovule, etc. Thecomposition may also be applied to a vaginal insert, tampon, wipe orpad, and then administered to the vagina. Formulations may comprise afirst saccharide and a second saccharide or an organic acid, a solventand optionally a dermatologically acceptable carrier. As used herein,“dermatologically acceptable carrier” generally refers to a carrier thatis suitable for topical application to the keratinous tissue and iscompatible with a prebiotic. The dermatologically acceptable carrier maybe in a wide variety of forms such as, for example, simple solutions(water-based or oil-based), solid forms (e.g. gels or sticks) andemulsions.

Solvents may be either aqueous or non-aqueous. Water is a particularlypreferred aqueous solvent. Non-aqueous solvents may include, forexample, glycols, such as propylene glycol, butylene glycol, triethyleneglycol, hexylene glycol, polyethylene glycols, ethoxydiglycol, anddipropyleneglycol; alcohols, such as ethanol, n-propanol, andisopropanol; triglycerides; ethyl acetate; acetone; triacetin; andcombinations thereof. Typically, the solvent constitutes greater thanabout 75 wt/vol %, more preferably greater than about 85 wt/vol %, andstill more preferably greater than about 90 wt/vol %.

The compositions of the present invention are generally acidic, i.e.,have a pH less than about 7.0 and more preferably less than about 6.0,such as from about 3.0 to about 6.0 and still more preferably from about4.0 to about 5.0. In a particularly preferred embodiment the pH may bemaintained at a mildly acidic level to correspond to normal vaginalconditions. For example, the pH may be within a range of from about 3.0to about 6.0, in some embodiments from about 3.5 to about 5.0, and insome embodiments, from about 4.0 to about 4.5. The foregoing acid pH mayalso provide other benefits. For instance, when the composition isconfigured to form a gel, such as described below, a low pH level mayalso improve the gelation rate and gel strength to reduce the likelihoodof leakage just after insertion of the composition into the vagina.

In view of the foregoing, in certain embodiments the composition maycomprise a first therapeutic agent consisting of arabinose or2-deoxy-D-ribose and a second therapeutic agent consisting of trehalose,lactulose, pectin, α-cyclodextrin or malic acid, wherein the totalamount of therapeutic agent is from about 0.1 to about 2.0 wt/vol % andthe ratio of the first therapeutic agent to the second therapeutic agentmay range from about 1:1 to about 1:50, such as from about 1:1 to about1:20 and more preferably from about 1:1 to about 1:5. In otherembodiments the composition may have a pH from about 3.0 to about 6.0,more preferably from about 3.5 to about 5.0, and comprise firsttherapeutic agent consisting of trehalose or lactulose and a secondtherapeutic agent consisting of arabinose, 2-deoxy-D-ribose, pectin,α-cyclodextrin or malic acid, wherein the total amount of therapeuticagent is from about 0.1 to about 2 wt/vol % and the ratio of the firsttherapeutic agent to the second therapeutic agent may range from about1:1 to about 1:50, such as from about 1:1 to about 1:20 and morepreferably from about 1:1 to about 1:5.

In one particular embodiment of the present invention, for example, thecomposition is configured to rapidly form a gel when applied to thevagina. A “gel” is a colloid in which a disperse phase combines with adispersion medium to produce a jelly-like, solid or semi-solid material.The gel may form in less than about one hour, in some embodiments lessthan about one minute, and in some embodiments, less than about 30seconds. Among other things, such rapid gelation reduces the likelihoodof leakage during use. In addition, because the gel may formintravaginally, it is more likely to retain its structure and shape overan extended period of time. In this manner, the gel may provide theprolonged release of a therapeutic agent that inhibits and/or treatsvaginal infection. For instance, the gel may remain within the vaginafor about 2 to about 48 hours to provide the desired effect.

Although a variety of compounds may be employed, water is usuallyemployed as the dispersion medium for the gel to optimizebiocompatibility. Other possible dispersion mediums include non-aqueoussolvents, including glycols, such as propylene glycol, butylene glycol,triethylene glycol, hexylene glycol, polyethylene glycols,ethoxydiglycol, and dipropyleneglycol; alcohols, such as ethanol,n-propanol, and isopropanol; triglycerides; ethyl acetate; acetone;triacetin; and combinations thereof. Typically, the dispersion medium(e.g., water) constitutes greater than about 75 wt/vol %, in someembodiments greater than about 90 wt/vol %, and in some embodiments,from about 95 to about 99 wt/vol % of the composition.

The disperse phase of the gel may be formed from any of a variety ofdifferent gelling agents, including temperature responsive(“thermogelling”) compounds, ion responsive compounds, and so forth.Thermogelling systems, for instance, respond to a change in temperature(e.g., increase in temperature) by changing from a liquid to a gel.Generally speaking, the temperature range of interest is from about 25°C. to about 40° C., in some embodiments from about 35° C. to about 39°C., and in one particular embodiment, at the human body temperature(about 37° C.). Compositions that change state at about this temperatureare useful because they will remain in a body cavity, for example, afterthey have been delivered. Any of a variety of thermogelling compoundsthat are capable of gelling when applied to the vagina may be used inthe present invention. In some cases, thermogelling block copolymers,graft copolymers, and/or homopolymers may be employed. For example,polyoxyalkylene block copolymers may be used in some embodiments of thepresent invention to form a thermo-gelling composition. Suitablethermo-gelling compositions may include, for example, homopolymers, suchas poly(N-methyl-N-n-propylacrylamide), poly(N-n-propylacrylamide),poly(N-methyl-N-isopropylacrylamide), poly(N-n-propylmethacrylamide),poly(N-isopropylacrylamide), poly(N,n-diethylacrylamide);poly(N-isopropylmethacrylamide), poly(N-cyclopropylacrylamide),poly(N-ethylmethyacrylamide), poly(N-methyl-N-ethylacrylamide),poly(N-cyclopropylmethacrylamide), and poly(N-ethylacrylamide). Stillother examples of suitable thermogelling polymers may include celluloseether derivatives, such as hydroxypropyl cellulose, methyl cellulose,hydroxypropylmethyl cellulose, and ethylhydroxyethyl cellulose. Moreoverthermogelling polymers may be made by preparing copolymers between(among) monomers, or by combining such homopolymers with otherwater-soluble polymers, such as acrylic monomers (e.g., acrylic ormethacrylic acid, acrylate or methacrylate, acrylamide ormethacrylamide, and derivatives thereof).

The compositions of the present invention may also include an ionresponsive compound. Such compounds are generally well known in the art,and tend to form a gel in the presence of certain ions or at a certainpH. For instance, one suitable class of ion responsive compounds thatmay be employed in the present invention is anionic polysaccharides.Anionic polysaccharides may form a three-dimensional polymer networkthat functions as the disperse phase of the gel. Generally speaking,anionic polysaccharides include polysaccharides having an overallanionic charge, as well as neutral polysaccharides that contain anionicfunctional groups.

Any of a variety of anionic polysaccharides capable of forming a gelwhen contacted with vaginal mucosa may be used in the present invention.Such gel-forming anionic polysaccharides are typically stable over thenormal acidic pH values found in the vagina (e.g., from about 2.5 toabout 5.5). For instance, some suitable examples of gel-forming anionicpolysaccharides include natural gums, such as gellan gum and alginategums (e.g., ammonium and alkali metal of salts of alginic acid);chitosan; carboxymethylcellulose, pectins, carrageenan, xantham gum, andderivatives or salts thereof. The particular type of anionicpolysaccharide selected will depend, in part, on the nature of thecomposition and the other components used therein. For example,carrageenan is sensitive to particular types of cations, e.g., ittypically gels in the presence of potassium but not sodium. Glycuronans,likewise, typically gel in the presence of divalent cations (e.g.,Ca2+), but not monovalent cations (e.g., Na+). Xanthan gum may gel inthe presence of divalent cations, but only at a relatively high pH.

Although any of the above-described anionic polysaccharides may be usedin the present invention, gellan gum is particularly desired for use inthe present invention, either alone or in combination with other gellingagents, because it is able to form a gel in the presence of a widevariety of different cations, including both monovalent and divalentcations. Gellan gum is intended to encompass any form of gellan,including native gellan, clarified gellan, deacylated gellan,nonacylated gellan (e.g., produced from genetically engineeredbacteria), clarified gellan (the polysaccharide is fully or partiallyremoved from the bacterial debris), chemically modified gellan, etc.Various types of gellan gums and methods for forming such gums aredescribed in U.S. Pat. Nos. 4,326,052; 4,326,053, 4,377,636; 4,385,123,and 4,563,366. Suitable gellan gums are commercially available from avariety of different sources. For example, GELRITE™ gellan gum isavailable from Sigma-Aldrich Chemical Co. of St. Louis, Mo., and isproduced from a naturally occurring polysaccharide after deacylation andclarification. Deacylated gellan is also available from CP Kelco U.S.,Inc. of Chicago, Ill. under the name KELCOGEL®.

Gellan gum may be either high or low acyl gellan. In the high acyl (or“native”) form, two acyl substituents, acetate and glycerate, arepresent. Both substituents are located on the same glucose residue and,on average, there is one glycerate per repeat unit and one acetate perevery two repeat units. In the low acyl form, the acyl groups may bewholly or partially removed through deacylation. The degree ofdeacylation of deacylated gellan gums may be at least about 20%, in someembodiments at least about 50%, and in some embodiments, at least about75%. Alternatively, the low acyl gellan gum may simply be “nonacylated”in that it is formed without acyl groups by genetically engineeredbacteria. Regardless of the manner in which they are formed, low acylgellan gums generally have a gelation temperature within the range 30 to50° C., which may be particularly well suited for use in the presentinvention so that it may gel at body temperatures of about 37° C., butremain stable at typical storage and transportation temperatures ofabout 25° C. In addition, low acyl gellan gums are also firm andelastic, and thus may retain their shape after delivery to the vaginalcavity.

In most embodiments the gelling agent(s) are present in an amount offrom about 0.01 to about 10.0 wt/vol %, in some embodiments from about0.05 to about 5.0 wt/vol %, and in some embodiments, from about 0.1 toabout 1.0 wt/vol % of the composition.

If desired, a gelling composition may be provided in any desired form(e.g., liquid, powder, etc.). In fact, one particular benefit of thecomposition is that it may be administered as a liquid, which allows forthe selection of a wider variety of administration techniques than wouldotherwise be available for a solid or semi-solid gel. One technique thatmay be employed includes dispensing the composition through a liquidapplicator, such as a syringe or tube, into the vaginal cavity. Theadministered volume of the composition may constitute a single dose ortwo or more doses. Although not necessarily required, the composition ofmay also be sterilized prior to administration. Sterilization may beaccomplished by any technique known in the art, such as using a gas(e.g., ethylene oxide), radiation (e.g., gamma), or heat (autoclaving).If desired, the composition may be subjected to one or more filtrationsteps prior to sterilization to help remove contaminants.

The urogenital compositions of the present invention may be applied to asuitable substrate, which in-turn may be used to apply the prebiotic toa user. Suitable applicators include a web, such as a wet laid tissueweb or air laid web, gauze, cotton swab, transdermal patch, container orholder. Particularly preferred applicators include fibrous webs,including flushable and non-flushable cellulosic webs and nonwoven websof synthetic fibrous material. Useful webs may be wet laid, air laid,meltblown, or spunbonded. Suitable synthetic fibrous material includesmeltblown polyethylene, polypropylene, copolymers of polyethylene andpolypropylene, bicomponent fibers including polyethylene orpolypropylene, and the like. Useful nonwoven webs may be meltblown,coform, spunbond, airlaid, hydroentangled nonwovens, spunlace, bondedcarded webs.

In certain embodiments, particularly those in which the urogenitalcomposition is applied to a web, it may be desirable that theformulation provide certain physical attributes, such as having asmooth, lubricious, non-greasy feel; the ability to at least partiallytransfer from the web to the user's skin; the capability to be retainedon the web at about room temperature; or the ability to be compatiblewith the web manufacturing process. In certain embodiments it ispreferred that at least a portion of the composition is transferred fromthe tissue to the user's skin in use.

The composition may be applied to a web during formation of the web orafter the web has been formed and dried, often referred to as off-lineor post-treatment. Suitable methods of applying the composition to a webinclude methods known in the art such as gravure printing, flexographicprinting, spraying, WEKO™, slot die coating, or electrostatic spraying.One particularly preferred method of off-line application is rotogravureprinting.

In those instances where the composition is added to the web duringformation of the web and prior to drying, it may be preferred to employan application method that incorporates the composition on the surfaceof the web. One method of adding the prebiotic to the web surface is byapplying the composition during creping of the tissue web. Surprisingly,the composition itself may be used as a creping composition or may becombined with other well-known creping compositions to apply thecomposition to a tissue web without significantly degrading importantweb properties such as strength, stiffness or sloughing.

Fibrous webs comprising a composition made according to the presentdisclosure can be incorporated into multi-ply products. For instance, inone aspect, a fibrous web made according to the present disclosure canbe attached to one or more other fibrous webs to form a wiping producthaving desired characteristics. The other webs laminated to the fibrousweb of the present disclosure can be, for instance, a wet-creped web, acalendered web, an embossed web, a through-air dried web, a crepedthrough-air dried web, an uncreped through-air dried web, an airlaidweb, and the like, and may or may not comprise a prebiotic.

In other embodiments, the composition could be applied to skin topromote, maintain, or enhance the balance of a healthy microflora.Application could be as a wipe, lotion, lubricant, cream, moisturizer,patch, or other topical application methods.

In certain embodiments the composition could be ingested to promote,maintain, or enhance the balance of a healthy microflora in thegastrointestinal tract.

TEST METHODS Therapeutic Effect Protocol

Colonies of L. crispatus and E. coli were prepared as follows. A colonyof L. crispatus was transferred to 7 ml MRS broth and incubatedanaerobically (using BD GasPak EZ anaerobe container system withindicator) at 37° C. without shaking for 18-20 hours. A colony of E.coli was transferred to 5 ml TSB broth and incubated aerobically(shaking at 100 rpm) at 37° C. for 18-20 hours.

Colonies were then inoculated as follows. Bacterial cultures were gentlyvortexed and 1 mL of each culture was transferred to a corresponding 2.0mL micro-centrifuge tube and then centrifuged for two minutes at 14,500rpm. The cultural supernatant was removed and the cell pellet wasre-suspended in 1 mL of 0.95% (wt/vol %) saline. The re-suspended colonywas then centrifuged for two minutes at 14,500 rpms and the supernatantwas removed. For L. crispatus, the pellet was re-suspended in 1 mL of0.95% saline to achieve ˜10⁷-10⁸ cfu/mL. For E. coli the pellet wasre-suspended in 1 mL of 0.95% saline to achieve ˜10⁸-10⁹ cfu/mL.

Media were prepared as follows.

TABLE 1 Ingredients g/L Peptone 15 Tryptone 10 Yeast Extract 10 Tween 801All of the ingredients in Table 1 were combined and the pH was adjustedto 6.5. The media were then autoclaved for 20 minutes at 125° C. Toevaluate the effect of various therapeutic agents on the growth ofbacteria the LAPT-g media, prepared as described above, was supplementedwith various therapeutic agents to a final test concentration between0.1 and 1.0%.

Five milliliters (5 mL) of each medium was transferred to a test tube induplicates for subsequent inoculation. A master mixture of L. crispatusand E. coli, prepared as described above, was prepared in 1,000:1 ratio.Each (5 ml) test tube was inoculated with the master mixture to give10⁵-10⁶ total CFU L. crispatus and 100-1,000 total CFU E. coli per tube.

To establish a negative control, one tube was vortexed and 100 μL wasremoved to determine the initial cell concentrations by serial dilutionsand plating (2 plates per dilution). L. crispatus is selected on MRSagar plates incubated anaerobically at 37° C. for two days. E. coli isselected on TSA plates incubated aerobically at 37° C. for one day. Theco-cultures were incubated in an anaerobic container with BD GasPaks at37° C. for 36 hours.

The effect of the carbon source on the ratio of L. crispatus to E. coliwas measured 36 hours after inoculation. The co-culture tube wasvortexed and 100 μL was removed to determine the final cellconcentrations by serial dilutions and plating (2 plates per dilution).L. crispatus is selected on MRS agar plates incubated anaerobically at37° C. for two days. E. coli is selected on TSA plates incubatedaerobically at 37° C. for one day.

EXAMPLES

Inventive samples were prepared by supplementing LAPT-g media, preparedas described above, with various therapeutic agents as described inTable 2, below. The therapeutic effect of the formulation was thenmeasured using the assay described above in the Test Methods sectionabove. The therapeutic effect is summarized below as the ratio of L.crispatus to E. coli. The combinations of therapeutic agents thatresulted in a ratio of L. crispatus/E. coli greater than the sum of theratios of the individual therapeutic agents were considered synergistic.

TABLE 2 Sample L. crispatus/ No. Therapeutic Agent (w/vol %) CAS # E.coli ratio 1 D-Trehalose (0.5%) + Pectin (0.5%) 6138-23-4, 9000-69-5653.3 2 D-Trehalose (0.5%) + α-Cyclodextrin (0.5%) 6138-23-4, 10016-20-3100.4 3 D-Trehalose (0.5%) + D-arabinose (0.5%) 6138-23-4, 28697-53-2197.3 4 Pectin (0.5%) + α-Cyclodextrin (0.5%) 9000-69-5, 10016-20-3 43.25 Pectin (0.5%) + α-Methyl-D-Glucoside (0.5%) 9000-69-5, 97-30-3 32.3 62-Deoxy-D-Ribose (0.1%) + Malic acid (0.1%) 533-67-5, 6915-15-7 340 72-Deoxy-D-Ribose (0.1%) + D-Trehalose (0.9%) 533-67-5, 6138-23-4 103.8 82-Deoxy-D-Ribose (0.1%) + Lactulose (0.1%) 533-67-5, 4618-18-2 562.3 9D-Trehalose (0.1%) + Pectin (0.4%) 6138-23-4, 9000-69-5 315.7 10 Dextrin(0.1%) + Pectin (0.4%) 9004-53-9, 9000-69-5 245.6 11 Dextrin (0.1%) +Pectin (0.4%) 9004-53-9, 9000-69-5 184.4 12 D-Trehalose (0.5%) + P95 FOS(0.5%) 6138-23-4, 9005-80-5 163.7 13 D-Trehalose (0.4%) + HSI Inulin(0.6%) 6138-23-4, 9005-80-5 126.5 14 D-Trehalose (0.8%) + HSI Inulin(0.2%) 6138-23-4, 9005-80-5 109.3 15 Malic acid (0.1%) + Pectin (0.4%)6915-15-7, 9000-69-5 105.5 16 Pectin (0.5%) + HSI Inulin (0.5%)9000-69-5, 9005-80-5 105.1 17 Salicin (0.1%) + Pectin (0.4%) 138-52-3,9000-69-5 81.3 18 Pectin (0.5%) + P95 FOS (0.5%) 9000-69-5, 9005-80-565.8 19 D-Trehalose (0.5%) + HSI Inulin (0.5%) 6138-23-4, 9005-80-5 40.520 α-Cyclodextrin (0.5%) + α-Methyl-D-Glucoside (0.5%) 10016-20-3,97-30-3 33.9 21 2-Deoxy-D-Ribose (0.1%) 533-67-5 1.7 22α-Methyl-D-Glucoside (1%) 97-30-3 3 23 α-Cyclodextrin (1%) 10016-20-36.5 24 D-Arabinose (1%) 28697-53-2 2.2 25 Dextrin (0.1%) 9004-53-9 0.926 D-Trehalose (1%) 6138-23-4 30.1 27 HSI Inulin (1%) 9005-80-5 0.5 28Lactulose (0.1%) 4618-18-2 3.9 29 Malic acid (0.1%) 6915-15-7 30.7 30P95 FOS (1%) 9005-80-5 1.1 31 Pectin (0.5%) 9000-69-5 9.5 32 Salicin(0.1%) 138-52-3 1.5

In view of the foregoing description and examples, the present inventionprovides, in a first embodiment, a composition comprising a firsttherapeutic agent consisting of a pentose or a disaccharide and a secondtherapeutic agent selected from the group consisting of a non-digestiblepolysaccharide, an organic acid, a cyclodextrin, a pectic substance, apentose or a disaccharide wherein first therapeutic and the secondtherapeutic agent are different.

The composition of the first embodiment may be formulated as a liquid, asolution, a paste or a gel.

The invention further provides in a second embodiment the composition ofthe first embodiment wherein the weight ratio of the first therapeuticand the second therapeutic agent is from about 1:1 to about 1:50.

Still further, the invention provides in a third embodiment thecomposition of the first or second embodiment, wherein the firsttherapeutic comprises from about 0.1 to about 2.0 wt/vol %, the secondtherapeutic agent comprises from about 0.1 to about 2.0 wt/vol % and thepH of the composition is from about 3.5 to about 6.0.

The invention also provides in a fourth embodiment the composition ofany one of the first through the third embodiments where the firsttherapeutic agent is a pentose selected from the group consisting ofribose, ribulose, arabinose, xylose, xylulose, and lyxose, methylβ-D-ribofuranoside and 2-deoxy-D-ribose.

Additionally, the invention provides in a fifth embodiment thecomposition of any one of the first through the fourth embodiments wherethe first therapeutic agent is a disaccharide selected from the groupconsisting of lactulose, trehalose, rhamnose maltose, maltotriose,lactose and lactitol.

The invention further provides in a sixth embodiment the composition ofany one of the first through the fifth embodiments where the secondtherapeutic agent is an organic acid selected from the group consistingof citric acid, lactic acid, methyllactic acid, phenyllactic acid, malicacid, mandelic acid, glycolic acid, tartronic acid, tartaric acid andgluconic acid.

The invention also provides in a seventh embodiment the composition ofany one of the first through the sixth embodiments where the secondtherapeutic agent is a non-digestible polysaccharide selected from thegroup consisting of dextrin, inulin, fructo-oligosaccharide (FOS) andisomalto-oligosaccharides.

The invention further provides in an eighth embodiment the compositionof any one of the first through the seventh embodiments where the firsttherapeutic agent is a pentose or a disaccharide and the secondtherapeutic agent is selected from the group consisting of pectin,α-cyclodextrin, dextrin, inulin and malic acid.

The invention also provides in a ninth embodiment the composition of anyone of the first through the eighth embodiments wherein the compositionsynergistically promotes the growth of L. crispatus relative to E. colisuch that the therapeutic effect is greater than about 30 and morepreferably greater than about 50 and still more preferably greater thanabout 100.

The invention further provides in a tenth embodiment a compositioncomprising a first therapeutic agent consisting of a non-digestiblepolysaccharide and a second therapeutic agent selected from the groupconsisting of an organic acid, a cyclodextrin, a pectic substance, apentose or a disaccharide.

The composition of the tenth embodiment may be formulated as a liquid,paste or a gel having a pH from about 3.0 to about 5.0 and comprisingfrom about 0.1 to about 2.0 wt/vol % of the first therapeutic agent andfrom about 0.1 to about 2.0 wt/vol % of the second therapeutic agent.

In an eleventh embodiment the present invention provides a compositioncomprising a first therapeutic agent selected from the group consistingof α-methyl-d-glucoside, 3-methyl-D-glucopyranoside and salicin and asecond therapeutic agent selected from the group consisting of apentose, a disaccharide, a non-digestible polysaccharide, an organicacid, a cyclodextrin and a pectic substance. In a particularly preferredembodiment the first therapeutic agent is α-methyl-d-glucoside orsalicin and the second therapeutic agent is pectin or α-cyclodextrin.

The invention also provides in an twelfth embodiment a method formaintaining a healthy microflora balance in the urogenital area, themethod comprising topically administering to the urogenital area of apatient in need thereof the composition of any one of the first throughthe eleventh embodiments.

In still another embodiment the present invention provides a method forenhancing lactobacillus growth or activity in vivo comprisingadministering a composition of any one of the first through the eleventhembodiments to a patient in need thereof.

What is claimed is:
 1. A composition comprising a first therapeuticagent consisting of a pentose or a disaccharide and a second therapeuticagent selected from the group consisting of a non-digestiblepolysaccharide, an organic acid, a cyclodextrin, a pectic substance, apentose or a disaccharide wherein first therapeutic and the secondtherapeutic agent are different.
 2. The composition of claim 1 whereinthe weight ratio of the first therapeutic and the second therapeuticagent is from about 1:1 to about 1:50.
 3. The composition of claim 1wherein the first therapeutic agent comprises from about 0.1 to about2.0 wt/vol %, the second therapeutic agent comprises from about 0.1 toabout 2.0 wt/vol %.
 4. The composition of claim 1 wherein the firsttherapeutic agent is a pentose selected from the group consisting ofribose, ribulose, arabinose, xylose, xylulose, and lyxose, methylβ-D-ribofuranoside and 2-deoxy-D-ribose.
 5. The composition of claim 1wherein the first therapeutic agent is a disaccharide selected from thegroup consisting of lactulose, trehalose, rhamnose maltose, maltotriose,lactose and lactitol.
 6. The composition of claim 1 wherein the secondtherapeutic agent is an organic acid selected from the group consistingof citric acid, lactic acid, methyllactic acid, phenyllactic acid, malicacid, mandelic acid, glycolic acid, tartronic acid, tartaric acid andgluconic acid.
 7. The composition of claim 1 wherein the secondtherapeutic agent is a non-digestible polysaccharide selected from thegroup consisting of dextrin, inulin, fructo-oligosaccharide (FOS),lactulose and isomalto-oligosaccharides.
 8. The composition of claim 1wherein the second therapeutic agent is selected from the groupconsisting of pectin, α-cyclodextrin, dextrin, inulin and malic acid. 9.The composition of claim 1 wherein the composition synergisticallypromotes the growth of L. crispatus relative to E. coli such that thetherapeutic effect is greater than about
 30. 10. A compositioncomprising a first therapeutic agent consisting of a cyclodextrin or apectic substance and a second therapeutic agent selected from the groupconsisting of a pectic substance, a pentose or a disaccharide, anon-digestible and a polysaccharide, wherein the first and secondtherapeutic agents are different.
 11. The composition of claim 10wherein the weight ratio of the first therapeutic agent and the secondtherapeutic agent is from about 1:1 to about 1:50.
 12. The compositionof claim 10 wherein the first therapeutic agent comprises from about 0.1to about 2.0 wt/v %, the second therapeutic agent comprises from about0.1 to about 2.0 wt/v %.
 13. The composition of claim 10 wherein thesecond therapeutic agent is a pentose selected from the group consistingof ribose, ribulose, arabinose, xylose, xylulose, and lyxose, methylβ-D-ribofuranoside and 2-deoxy-D-ribose.
 14. The composition of claim 10wherein the second therapeutic agent is a disaccharide selected from thegroup consisting of lactulose, trehalose, rhamnose maltose, maltotriose,lactose and lactitol.
 15. The composition of claim 10 wherein the firsttherapeutic agent is dextrin, inulin, or fructo-oligosaccharide (FOS)and the second therapeutic agent is 2-deoxy-D-ribose, trehalose,lactulose, arbinose, pectin, or α-cyclodextrin.
 16. The composition ofclaim 10 wherein the first therapeutic agent is a pectic substanceselected from the group consisting of pectin, pectate andpolygalacturonic acid and the second therapeutic agent isα-cyclodextrin.
 17. The composition of claim 10 wherein the compositionsynergistically promotes the growth of L. crispatus relative to E. colisuch that the therapeutic effect is greater than about
 30. 18. A methodfor maintaining a healthy microflora balance in the urogenital area of apatient in need thereof, the method comprising topically administeringto the urogenital area of a patient a composition comprising a firsttherapeutic agent consisting of a pentose or a disaccharide and a secondtherapeutic agent selected from the group consisting of a non-digestiblepolysaccharide, an organic acid, a cyclodextrin, a pectic substance, apentose or a disaccharide wherein the first therapeutic agent and thesecond therapeutic agent are different.
 19. The method of claim 18wherein administration of the composition increases lactobacillus growthor activity in vivo.
 20. The method of claim 18 wherein the firsttherapeutic agent is 2-deoxy-D-ribose, trehalose, lactulose or arbinoseand the second therapeutic agent is, dextrin, inulin, orfructo-oligosaccharide (FOS), pectin, α-cyclodextrin or malic acid, thefirst therapeutic agent being present in an amount from about 0.1 toabout 2.0 wt/vol % and the second therapeutic agent being present in anamount from about 0.1 to about 2.0 wt/vol %.